Random Substitutions

In a controlled randomization library, random nucleotides within a defined region will be mutated at a defined average mutation rate. GeneArt™ Controlled Randomization technology allows you to accurately direct the effective frequency of random nucleotide substitutions. And unlike error-prone PCR, you decide exactly which part(s) of the gene are affected—randomize the entire open reading frame or confine the mutagenesis to specific regions. Randomized constructs are rigorously controlled to help ensure that the unchanged portions of the sequence are unaffected and that randomized portions contain the distribution of nucleotides you require to encode the amino acid composition you’re looking for.

Precise selection of the average mutation rate (number of changes per construct)

Random mutagenesis throughout construct, or limited to defined positions

Transition/transversion ratio of 1:2

Fewer silent mutations

Low ancillary mutation rate

Fast production times

All GeneArt™ Controlled Randomization products are sequenced and subjected to statistical analysis to help ensure that they meet the following quality benchmarks:

Sequencing of up to 96 individual transformants to verify that customer’s specifications regarding nucleotide content and amino acid encoding have been met, and that sequence integrity of unmutated portions of the construct meets minimum requirements

Bulk sequencing to verify that nondegenerated portions of the construct have the correct sequence and randomized portions meet customer’s specifications and maintain an open reading frame

Real-time PCR prior to amplification to verify that library diversity objectives have been met